Correlation of histology and alpha1-fetoprotein resurgence in rat liver regeneration after experimental injury by galactosamine

Summary Intraperitoneal injections of galactosamine-HCl in rats were followed by transitory elevations of serum alpha-fetoprotein (AFP) concentrations. These were associated with regeneration of the damaged liver; a maximum of serum AFP was reached on day 4 (2.08±0.67 g/ml). In sera of untreated rats, serum AFP levels were <0.1 g/ml and no cellular AFP was detected in liver sections. Two days after galactosamine injections, AFP was localized for the first time in the cytoplasm of epithelial cells of bile ducts and canaliculi in portal spaces. The intensity of AFP staining reached a maximum between days 3 and 4. In addition, faint but distinct AFP-positive reactions were seen in the cytoplasm of randomly distributed hepatocytes. After day 5, AFP-staining cells rapidly disappeared. A strong correlation was noted between reappearance of AFP in sera, intensity of epithelial bile duct proliferation and cellular AFP staining.The present paper on alpha₁-fetoprotein is dedicated to Prof. K.E. Scheer, Institut für Nuklearmedizin/D.K.F.Z. Heidelberg, on the occasion of his 60th birthdaySupported by the Deutsche Forschungsgemeinschaft (Ku 257/3) D-5300 Bonn, Federal Republic of Germany     

冷缺血损伤后大鼠肝脏移植后肝脏再生的分子机制

目的： 研究不同冷缺血损伤条件下，大鼠肝脏移植后肝脏再生的分子机制。方法: 建立大鼠原位肝移植模型。实验分为: 冷缺血1 h、8 h和16 h组。观察各组的生存率，并在术后90 min、1、2、4和7 d收集标本，观察白介素-6(IL-6)、肿瘤坏死因子（TNF-α）、信号转导激活蛋白3 (STAT3)等表达情况。免疫组化检测细胞周期素D1的表达和肝细胞摄取溴脱氧尿核苷情况。比较实验各组肝脏移植后TNF-α和IL-6的表达水平。分析实验各组移植术后48 h 溴脱氧尿核苷染色阳性的肝细胞数。结果: 冷缺血1 h、8 h和16 h后肝脏移植物的存活率均为100％（>14 d）。移植术后90 min，与冷缺血1 h组相比，冷缺血8 h和16 h组大鼠移植肝内TNF-α等因子表达明显增加（P<0.05）。移植术后90 min，与冷缺血1 h和8 h组相比，16 h组IL-6的表达亦明显增强（P<0.05）。冷缺血8 h和16 h组STAT3活性明显增强。冷缺血8 h组细胞周期素D1在胞浆和细胞核内均有表达。冷缺血16 h组细胞周期素D1仅在核内表达。肝细胞复制活跃。与冷缺血1 h和8 h组相比，冷缺血16 h组术后48 h溴脱氧尿核苷染色阳性的肝细胞计数也明显增多（P<0.05）。结论: 大鼠肝脏移植物经受16 h冷缺血损伤后仍然能够启动，完成肝脏再生，修复组织损伤。大鼠肝脏移植后的肝脏再生可能通过TNF-α/IL-6／STAT3/cyclin D1/DNA合成的途径调节。     

大鼠肝纤维化过程中肝组织及血清中粗纤维调节素的动态变化研究

目的：观察粗纤维调节素（Un）在大鼠肝纤维化肝组织及血清中的动态变化，探讨其在正常和纤维化大鼠肝组织中的表达及其血清学检测的价值。方法：以CCl₄诱发大鼠肝纤维化模型，采用ABC免疫组化方法检测肝组织中Un分布，用ELISA方法检测血清中Un的含量。结果：随着CCl₄造模时间的延长，Un在肝组织中沉积增加，主要在汇管区、中央静脉壁及纤维间隔内分布。血清Un含量也随肝纤维化的加重而升高。结论：Un是肝组织中一种重要的细胞外基质成分，在肝纤维化过程中表达增加；血清Un可以作为反映肝纤维化的一种血清学指标。     

液压转基因技术应用于大鼠再生肝转基因实验

目的 探讨液压转基因技术(HDT)应用于大鼠再生肝转基因的条件和方法 . 方法 以2ml/s的速度将浓度为30mg/L的含目的 基因的质粒注射入大鼠尾静脉,于注射前/后不同时间进行大鼠2/3肝切除(PH),于PH后不同恢复时间称量大鼠体重(g)和再生肝重(g),计算肝系数(Lc),并从Lc±Lc*0%、*5%、*10%、*15%、*20%、*25%、*30%、*35%等15组中找出最佳组,作为计算不同恢复时间再生肝最适注射质粒溶液量的校正系数(Trc);取大鼠肝右叶中部组织制备冷冻切片,在波长488nm的荧光显微镜下观察、计数1万个细胞中的绿色荧光蛋白阳性细胞百分率. 结果 PH后注射生理盐水和注射空质粒对肝再生的影响与对照(只进行PH)相比无显著差异.PH前液压转基因的合适时间是PH前≥12h;PH后所有时间均可进行液压转基因.PH后对肝再生大鼠进行液压转基因的转基因溶液体积为大鼠体重(g)×9%×1/3×相应的校正系数(Trc).转入基因在体内的表达时间和丰度既受载体影响,又受插入的目的 基因影响. 结论 液压转基因技术亦可有效地应用于大鼠再生肝转基因研究.     

Intra-aortic balloon: evaluation of heparin-coating under various experimental conditions.

In a calf model, heparin coated intra-aortic balloon (IAB) was compared with standard balloon. In group 1, 9 of each IAB type were set to the automatic mode for 15 min, 45 min and 6 hours respectively, while in group 2, 3 of each IAB type were left deflated during 20 minutes to simulate balloon dysfunction. At the end of the procedures, 3 samples of each IAB were analyzed with scanning electron microscopy (SEM) for surface deposits. Macroscopically, the 12/12 heparin coated IAB of both groups and the 9/9 standard IAB of group 1 were free of deposits, whereas the 3/3 standard IAB of group 2 exhibited clot deposits. SEM revealed deposit-free surfaces in the 36/36 heparin coated samples of both groups, while 14/27 standard samples of group 1 (p<0.01 when compared with heparin coated samples) and 8/9 standard samples of group 2 (p = 0.02, same comparison) disclosed blood cells and fibrin deposits. Morphometrically, the proportion of standard sample surfaces covered with deposits, estimated according to a score system (0% = 0; 0.1-25% = 1; 25.1-50% = 2; 50.1-75% = 3; 75. 1-100% = 4), was 0.69+/-0.82 in group 1 (p<0.01 when compared with heparin coated samples) and 1.22+/-0.83 in group 2 (p<0.01, same comparison). Thus heparin coated IAB presents no deposits either after 6 hours of intravascular ballooning or after 20 minutes of stagnation. It seems to be a promising strategy for patients with absolute or relative contraindications to systemic heparinization.     

Clinical implications of advances in liver regeneration

Remarkable advances have been made recently in the area of liver regeneration. Even though liver regeneration after liver resection has been widely researched, new clinical applications have provided a better understanding of the process. Hepatic damage induces a process of regeneration that rarely occurs in normal undamaged liver. Many studies have concentrated on the mechanism of hepatocyte regeneration following liver damage. High mortality is usual in patients with terminal liver failure. Patients die when the regenerative process is unable to balance loss due to liver damage. During disease progression, cellular adaptations take place and the organ microenvironment changes. Portal vein embolization and the associating liver partition and portal vein ligation for staged hepatectomy are relatively recent techniques exploiting the remarkable progress in understanding liver regeneration. Living donor liver transplantation is one of the most significant clinical outcomes of research on liver regeneration. Another major clinical field involving liver regeneration is cell therapy using adult stem cells. The aim of this article is to provide an outline of the clinical approaches being undertaken to examine regeneration in liver diseases.     

胰岛再生源蛋白与肝脏疾病和肝再生

胰岛再生源蛋白 (Reg) 是相对分子质量为16kD的分泌型蛋白,属于C类凝集素超家族成员。Reg蛋白是1个多功能分子,主要参与调控胰腺、胃、肠和肝脏中细胞的生长和增殖,在消化性疾病及癌症的发展和预后中发挥重要作用。我们主要综述了Reg对肝脏疾病和肝再生调控的研究进展,包含其在消化性疾病中介导的信号通路,为开展Reg在相关疾病诊疗及促进肝再生中的应用研究提供重要理论基础。     

索拉菲尼对肝硬化大鼠部分肝切除术后肝脏再生的影响

目的: 在大鼠肝硬化模型的基础上行肝脏部分切除(PH),研究索拉菲尼(sorafenib)对大鼠肝脏再生的影响。方法: 使用二乙基亚硝胺(DEN)诱导Wistar大鼠肝硬化,成功建立30只肝硬化大鼠PH模型后,随机分2组,每组15只。术后第1 d开始,分别给予实验组索拉菲尼(30 mg·kg^-1·d^-1)、对照组生理盐水灌胃10 d后处死。留取PH后及实验结束后的血液及肝脏标本,检测2组肝脏再生率(LRR),增殖细胞核抗原(PCNA),生化指标: 丙氨酸转移酶(ALT)和血清白蛋白(ALB)、血清总胆红素(TBIL)和血清直接胆红素(DBIL)的变化,血管生成相关因子:血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR-2)、血小板源性生长因子受体β(PDGFR-β),以及肝脏微血管密度(MVD)的变化。结果: (1)LRR在实验组及对照组分别为45.43%±3.36%和44.21%±2.77%,无显著差异(P>0.05);(2)免疫组织化学(IHC)没有检测到PCNA;(3)2组的生化指标无显著差异(P>0.05);(4)实验组VEGFR-2和PDGFR-β的表达受到抑制,MVD降低,并且实验组与对照组差异有统计学意义(P<0.01)。结论: 索拉菲尼虽然对肝硬化血管再生相关因子有抑制作用,但是对肝细胞再生和肝功能没有明显影响。     

大鼠肝再生过程中线粒体氧化磷酸化的调控

目的：探讨肝部分切除后肝再生过程中线粒体能量代谢的调控。方法：用雄性Ｗｉｓｔａｒ大鼠施行肝部分切除复制肝再生模型,肝部分切除后分别观察０５、１、２、４和７ｄ５个肝再生组以及５个相应的对照组,差速离心法分离肝线粒体并用氧电极极谱法测定其氧化磷酸化活性。结果：肝部分切除后肝再生过程中线粒体呼吸控制率（ＲＣＲ）明显高于相应对照组,其中肝部分切除后０５ｄ和４ｄ为二个峰值,７ｄ时降至对照组水平,早期ＲＣＲ的升高主要是Ｒ３升高的所致,１ｄ后ＲＣＲ升高是Ｒ４下降所致。磷氧比值（Ｐ／Ｏ）的变化类似于ＲＣＲ。结论：肝部分切除后肝再生过程中线粒体通过氧化磷酸化偶联增强来适应肝再生的能量需求,这种增强机制很可能主要是通过降低线粒体内膜通透性实现的。     

Alpha-fetoprotein synthesis in different lines of adult mice during regeneration of the liver

The concentration of alpha-fetoprotein (AFP) in the sera of adult mice of 12 different lines and F₁ hybrids of lines SWR and B10D2 was determined by radioimmunodiffusion in agar during regeneration of the liver after poisoning with CCl₄. In the mice of 6 of 10 lines differences in the AFP concentration between females and males were statistically significant. Interlinear differences also were found: The mean AFP concentrations in the sera of inbred C57BL/6 and B10D2 mice were significantly lower than the corresponding values for mice of most other lines. The F₁ hybrids were intermediate as regards their AFP concentration between the two parental lines. Small but statistically significant differences were found between groups of male F₁ hybrids in direct and reciprocal corsses. It is suggested that induction of AFP synthesis during regeneration of the liver in adult mice is under polygenic control.Laboratory of Immunochemistry and Diagnosis of Tumors, Oncologic Scientific Center, Moscow. Inbred Animals Group, Laboratory of Gnotobiology, N. F. Gamaleya Institute of Epidemiology and Microbiology, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 7, pp. 71–75, July, 1978.     

几种因素对大鼠再生肝摄取氨基酸能力的影响——离体肝灌流的实验研究

本实验选用大鼠肝部分切除后24小时的残余肝离体灌流,研究了四种因素对残余肝摄取氨基酸能力的影响。结果表明:1.肝部分切除后24小时,残余肝摄取氨基酸的量增加。肝脏摄取氨基酸能力的增强依赖于肝细胞蛋白质合成的增加。在一定范围内,肝脏摄取氨基酸的量随肝切除量的增加而增多。2.胰岛素能促进残余肝摄取氨基酸,高血糖素无此作用,但高血糖素与胰岛素联合施用则使残余肝摄取氨基酸的能力进一步增强。3.大鼠再生肝提取液中含有某种能促进残余肝摄取氨基酸的物质。     

#br# microRNAs在肝再生中的作用研究进展

目的 microRNAs是一类非编码蛋白的小RNA分子,通过与靶基因mRNA 3’端的非编码区（3’UTRS）结合而在转录后水平调控基因表达,调节细胞增殖、分化、代谢、凋亡、器官发育等生物学过程。肝脏有很强的再生能力,是研究再生的重要材料。因此,我们在文中总结了microRNAs对肝再生调节作用的研究进展。     

Long term liver blood flow measurements in rats under physiological conditions

Summary An improved method using the principle of internal calorimetry as an automatic recorder is described for long term studies measuring liver blood flow in 28 rats. The daily measurements of liver blood flow under physiological conditions showed very constant values with a low standard deviation. One problem encountered was the development of connective tissue around the thermocouple in three rats. Methods to avoid this complication are discussed. This method seems to be appropriate to study liver blood flow in the rat in long term physiological or pharmacological studies.     

大鼠肝再生氨基酸靶向代谢组学分析

目的 探讨氨基酸代谢对大鼠肝再生 (LR)的调控作用。 方法 大鼠随机分为10组,每组5只,包括9个部分肝切除组 (PH)和1个正常对照组。运用质谱选择反应检测扫描/多反应检测扫描 (SRM/MRM)对大鼠肝再生中10个时间点的20种氨基酸的含量进行靶向代谢组学鉴定,并对氨基酸含量变化进行聚类分析。 结果 丙氨酸在30、36和72 h上调;精氨酸在6和12 h上调;天冬氨酸在6、12和36 h上调;谷氨酸在6、12、30、36、72和120 h上调;组氨酸在12、24、30、36、72和120 h上调;谷氨酰胺在72 h上调;异亮氨酸在24 h下调。通过对氨基酸聚类分析发现,这些氨基酸可以聚为3类。 结论 肝再生过程中多种氨基酸发生了变化,且贯穿大鼠肝再生的全过程。氨基酸含量的变化,对肝再生中肝细胞增殖有重要作用。     

肝再生对大鼠胎肝细胞脾内移植后增殖的影响

目的:研究肝再生状态对大鼠胎肝细胞脾内移植后增殖影响。方法:分离孕3周SD大鼠胚胎肝细胞,将其移植入70%肝切除肝再生模型大鼠脾内,分别于移植后7 d和30 d应用流式细胞仪检测肝切除大鼠残肝细胞的细胞周期,用图像分析法检测脾内移植胎肝细胞面积密度。结果:移植后7 d,肝切除鼠残肝细胞S和G₂/M期细胞比例都明显少于对照组(P＜0.05),而其脾内移植胎肝细胞面积密度则显著高于对照组(P＜0.05);30 d后,各组间残肝细胞再生状态与移植胎肝细胞的面积密度均无明显差异。 结论:肝再生状态有利于大鼠胎肝细胞脾内移植后的增殖。     

大鼠肝再生中环状RNA表达谱的变化及其对自噬的影响

目的 探讨大鼠肝再生中环状RNA(circRNA)的表达变化以及对自噬的影响.方法 以60只大鼠2/3肝切除0 h、2 h、6 h、12 h、24 h、30 h、36 h、72 h、120 h 和 168 h 后的再生肝 circRNAs、微小 RNAs(miRNAs)、mRNA的高通量测序数据为基础,分析其在肝再生中...     

Substrate channelling in a creatine kinase system of rat skeletal muscle under various pH conditions

The aim of this study was to evaluate myofibrillar creatine kinase (CK) activity and to quantify the substrate channelling of ATP between CK and myosin ATPase under different pH conditions within the integrity of myofibrils. A pure myofibrillar fraction was prepared using differential centrifugation. The homogeneity of the preparation and the purity of the fraction were confirmed microscopically and by enzymatic assays for contaminant enzyme activities. The specific activity of myofibrillar CK reached 584 +/- 33 nmol PCr min(-1) mg(-1) at pH 6.75. Two methods were used to detect CK activity: (1) measurement of direct ATP production, and (2) measurement of PCr consumption. This method of evaluation has been tested in experiments with isolated creatine kinase. No discrepancy in CK activity between the methods was observed in the pH range tested (6.0-7.5). However, the same procedures resulted in a significant discrepancy between the amounts of reacted PCr and produced ATP within the pure myofibrillar fraction. This discrepancy represents the portion of ATP produced by the CK reaction, which is preferentially channelled to the myosin ATPase before diffusing into the bulk solution. The maximum evaluated difference reached 42.3 % at pH 6.95. The substrate channelling between myofibrillar-bound CK and myosin ATPase was evaluated under various pH levels within the physiological range and it reached a maximum value in a slightly acidic environment. These results suggest that ATP/ADP flux control by the CK system is more important at lower pH, corresponding to the physiological state of muscle fatigue.